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Quantitative Readout of Methionine Residue Solvent Accessibility in E. coli cells Using Radiolytic Hydroxyl Radical Labeling and Mass Spectrometry

ABSTRACT: Reactive oxygen species play a crucial role in cellular processes, but their effects on protein structure and function in vivo remain challenging to study. Here, we used synchrotron-based X-ray footprinting to probe protein structure in live E. coli, using quantitative LC-coupled mass spectrometry of methionine oxidation (MSox) with X-ray dose as a variable. A label-free proteomic analysis identified 2104 proteins from E. coli, with 465 proteins exhibiting MSox modifications distributed across multiple cellular compartments. Changes in MSox modification with increasing X-ray dose revealed a correlation between rates of modification and solvent-accessible surface area in vivo for selected proteins, providing a direct probe of protein structure and its conformational plasticity in the cell.

INSTRUMENT(S):

ORGANISM(S): Escherichia Coli

TISSUE(S): Cell Culture

SUBMITTER: Erik Farquhar

LAB HEAD: Mark R.

PROVIDER: PXD061660 | Pride | 2025-04-10

REPOSITORIES: Pride

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			Action	DRS
	BNL_021424_Replicate1_0.raw	Raw
	BNL_021424_Replicate1_20.raw	Raw
	BNL_021424_Replicate1_50.raw	Raw
	BNL_021424_Replicate2_0.raw	Raw
	BNL_021424_Replicate2_20.raw	Raw

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